confocal microscopy zeiss 710 uv/vis meta  (Carl Zeiss)

Journal: Cancer Management and Research

Article Title: Deguelin Restores Paclitaxel Sensitivity in Paclitaxel-Resistant Ovarian Cancer Cells via Inhibition of the EGFR Signaling Pathway

doi: 10.2147/CMAR.S457221

Figure Lengend Snippet: Cotreatment with deguelin and paclitaxel induced cell cytotoxicity in SKOV3-TR ovarian cancer cells. ( A ) SKOV3 (5 x 10 3 ) and SKOV3-TR (5 x 10 3 ) cells were seeded in 96-well plates and further incubated for 24 h. Cells were treated with serially diluted paclitaxel (0–1000 nM) for 48 h. ( B ) SKOV3-TR cells were treated with deguelin (0, 2.5, 5, 10, and 20 µM) for 48 h. ( C ) SKOV3-TR (5 x 10 3 ) cells were seeded in a 96-well plate for 24 h, then treated with serially diluted paclitaxel (0–500 nM) together with dose-different combinations (0, 5, and 10 µM) of deguelin. The cell viability was examined by WST-1 analysis. ( D ) SKOV3-TR (5 x 10 5 ) cells were seeded in 100 mm plates. After 24 h, cells were treated with DMSO (mock), paclitaxel (200 nM), deguelin (10 µM), and cotreated with deguelin (10 µM) and paclitaxel (200 nM) for 12 h, 24 h, and 48 h, respectively. ( E ) SKOV3-TR cells were treated with paclitaxel (200 nM) together with dose-different combinations (0, 1, 5, and 10 µM) of deguelin, and treated with dose-different combinations (0, 10, 100, and 200 nM) of paclitaxel together with deguelin (10 µM). At 48 h after treatment, cells were subjected to immunoblotting to examine the level of cleaved PARP. Actin was used as a loading control. ( F ) SKOV3-TR cells were treated with DMSO (mock), paclitaxel (200 nM), deguelin (10 µM), and treated with a combination of paclitaxel (200 nM) and deguelin (10 µM) for 48 h. The morphological changes of SKOV3-TR cells were observed by microscopy at 48 h. ( G ) SKOV3-TR (2 x 10 5 ) cells were seeded in 6-well plates and treated with paclitaxel (200 nM) and deguelin (10 µM) for 48 h. Living, early apoptotic, and late apoptotic cells were determined using Annexin V-FITC and PI staining, followed by flow cytometry. The statistical experiments were repeated three times. *p < 0.05 and ***p < 0.001 analyzed by concentration were considered significant.

Article Snippet: The cells were visualized by UV microscopy (Zeiss, Oberkochen, Germany).

Techniques: Incubation, Western Blot, Control, Microscopy, Staining, Flow Cytometry, Concentration Assay

Journal: Cancer Management and Research

Article Title: Deguelin Restores Paclitaxel Sensitivity in Paclitaxel-Resistant Ovarian Cancer Cells via Inhibition of the EGFR Signaling Pathway

doi: 10.2147/CMAR.S457221

Figure Lengend Snippet: The expression of P-glycoprotein (P-gp) and its efflux function were not affected by deguelin or paclitaxel in SKOV3-TR cells. ( A ) The expression levels of P-gp in SKOV3 and SKOV3-TR cells were confirmed by immunoblotting. Actin was used as a loading control. ( B ) SKOV3-TR cells were treated with paclitaxel, deguelin, and cotreated with paclitaxel and deguelin as indicated. At 48 h posttreatment, the expressions of P-gp were examined by immunoblotting. ( C ) SKOV3-TR cells were treated with paclitaxel and deguelin as above. At 48 h of treatment, cells were additionally treated with the fluorogenic P-gp substrate fluo-3/AM for 1 h. Fluorescent SKOV3-TR cells were visualized by UV microscopy. SKOV3 cells were also treated with fluo-3/AM as a positive control.

Article Snippet: The cells were visualized by UV microscopy (Zeiss, Oberkochen, Germany).

Techniques: Expressing, Western Blot, Control, Microscopy, Positive Control

Journal: Frontiers in Plant Science

Article Title: Endogenous cell wall degrading enzyme LytD is important for the biocontrol activity of Bacillus subtilis

doi: 10.3389/fpls.2024.1381018

Figure Lengend Snippet: Direct effect of B . subtilis GLB191 and its derivatives on P. viticola zoospores on grapevine leaves. B . subtilis GLB191(pHY300) [GLB191(pHY300)], Δ lytD (pHY300), and Δ lytD (pHY- lytD ) were cultured in PDB medium for 48 h at 37°C. Grapevine ( V. vinifera cv. Red Globe) leaves were treated with PDB medium (PDB) or the culture of GLB191(pHY300), Δ lytD (pHY300), and Δ lytD (pHY- lytD ), and inoculated with P. viticola sporangia 2 h post-treatment. Leaf discs were then punched out from leaves 24 h post-inoculation, and the number of infection sites (i.e., stomata with encysted zoospores of P. viticola ) was determined by UV epifluorescence observations after aniline blue staining of the pathogen. (A) Representative fields of fluorescence microscopy observations. Arrows indicate the infection sites (encysted zoospores). Scale bars represent 10 μm. (B) Values are the mean number of infection sites (means ± SD) for 9 discs from three replicates. Different letters on the column indicate significant different at p < 0.05 according to the Duncan test. The data are representative of three independent experiments.

Article Snippet: Three representative fields of each disc and four discs per leaf were observed by ultraviolet (UV) epifluorescence microscopy (Nikon Ti-E, Tokyo, Japan).

Techniques: Cell Culture, Infection, Staining, Fluorescence, Microscopy

Journal: Frontiers in Plant Science

Article Title: Endogenous cell wall degrading enzyme LytD is important for the biocontrol activity of Bacillus subtilis

doi: 10.3389/fpls.2024.1381018

Figure Lengend Snippet: Callose deposition induced by B . subtilis GLB191 and its derivatives on grapevine leaves. Grapevine ( V. vinifera cv. Red Globe) leaves were treated with PDB medium (PDB) or the culture of the wild-type B . subtilis GLB191(pHY300) [GLB191(pHY300)], Δ lytD (pHY300), and Δ lytD (pHY- lytD ). Callose production (fluorescent spots) was observed 3 days post-treatment by epifluorescence observations after aniline blue staining. (A) Representative fields observed using a fluorescence microscope. Scale bars represent 100 μm. (B) Values are the mean number of fluorescent spots (means ± SD) for 9 discs from three replicates. Different letters on the column indicate significant difference at p < 0.05 according to the Duncan test. The data are representative of three independent experiments.

Article Snippet: Three representative fields of each disc and four discs per leaf were observed by ultraviolet (UV) epifluorescence microscopy (Nikon Ti-E, Tokyo, Japan).

Techniques: Staining, Fluorescence, Microscopy